DOCSLIB.ORG

The Reliability of Estimating Ki Values for Direct, Reversible Inhibition Of

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
The Reliability of Estimating Ki Values for Direct, Reversible Inhibition Of Supplemental material to this article can be found at: http://dmd.aspetjournals.org/content/suppl/2015/09/09/dmd.115.066597.DC1 1521-009X/43/11/1744–1750$25.00 http://dx.doi.org/10.1124/dmd.115.066597 DRUG METABOLISM AND DISPOSITION Drug Metab Dispos 43:1744–1750, November 2015 Copyright ª 2015 by The American Society for Pharmacology and Experimental Therapeutics The Reliability of Estimating Ki Values for Direct, Reversible Inhibition of Cytochrome P450 Enzymes from Corresponding IC50 Values: A Retrospective Analysis of 343 Experiments s Lois J. Haupt, Faraz Kazmi, Brian W. Ogilvie, David B. Buckley, Brian D. Smith, Sarah Leatherman, Brandy Paris, Oliver Parkinson, and Andrew Parkinson XenoTech, LLC, Lenexa, Kansas (L.J.H., F.K., B.W.O., D.B.B., B.D.S., S.L.); and XPD Consulting, Shawnee, Kansas (B.P., O.P., A.P.) Received July 28, 2015; accepted September 8, 2015 ABSTRACT In the present study, we conducted a retrospective analysis of 343 in low (0.1 mg/ml or less) to maximize the unbound fraction of inhibitor. Downloaded from vitro experiments to ascertain whether observed (experimentally Under these conditions, predicted Ki values, based on IC50/2, corre- determined) values of Ki for reversible cytochrome P450 (P450) lated strongly with experimentally observed Ki determinations [r = inhibition could be reliably predicted by dividing the corresponding 0.940; average fold error (AFE) = 1.10]. Of the 343 predicted Ki values, IC50 values by two, based on the relationship (for competitive in- 316 (92%) were within a factor of 2 of the experimentally determined Ki hibition) in which Ki =IC50/2 when [S] (substrate concentration) = Km values, and only one value fell outside a 3-fold range. In the case of (Michaelis-Menten constant). Values of Ki and IC50 were determined noncompetitive inhibitors, Ki values predicted from IC50/2 values were dmd.aspetjournals.org under the following conditions: 1) the concentration of P450 marker overestimated by a factor of nearly 2 (AFE = 1.85; n = 13), which is to be substrate, [S], was equal to Km (for IC50 determinations) and spanned expected because, for noncompetitive inhibition, Ki =IC50 (not IC50/2). Km (for Ki determinations); 2) the substrate incubation time was short The results suggest that, under appropriate experimental conditions (5 minutes) to minimize metabolism-dependent inhibition and inhibitor with the substrate concentration equal to Km, values of Ki for direct, depletion; and 3) the concentration of human liver microsomes was reversible inhibition can be reliably estimated from values of IC50/2. at ASPET Journals on September 28, 2021 Introduction the P450 enzyme inhibited by the investigational drug (i.e., fractional When an investigational drug is evaluated in vitro as a direct, metabolism by an enzyme, or fm = 1.0) (Ito et al., 1998; Rodrigues reversible inhibitor of human liver cytochrome P450 (P450) enzymes et al., 2001). When other pathways contribute to clearance of the probe , based on the Food and Drug Administration’s (FDA’s) 2012 draft drug, such that fm 1, the fold increase in plasma AUC of the probe Guidance for Industry on drug interactions (http://www.fda.gov/ drug in the presence of the inhibitory drug is given by eq. 2: downloads/Drugs/GuidanceComplianceRegulatoryInformation/ 1 AUCR ¼ À Á ð2Þ Guidances/UCM292362.pdf), the need for a clinical drug interaction 1 × × þ 2 × ½I fm fm:enzyme 1 fm fm:enzyme 1þ study is based on eq. 1: Ki;unbound ½I A clinical drug interaction study is recommended for P450 en- AUCR ¼ R1 ¼ 1 þ ð1Þ Ki;unbound zymes other than CYP3A when the ratio R1 . 1.1 (where R1 5 1 1 [I]/Ki,unbound). The same criteria apply to significant circulating meta- where [I] is the maximum total (bound + unbound) plasma bolites, defined by the FDA as metabolites whose plasma AUC is $25% concentration of drug at steady state (Cmax,ss), and Ki,unbound is the of the parent AUC following dosing to steady state with the maximum dissociation constant for the enzyme-inhibitor complex for direct, clinical dose. The European Medicines Agency’s(EMA’s) 2012 reversible inhibition based on the concentration of unbound drug in the Guideline on the Investigation of Drug Interactions (http://www.ema. in vitro test system. AUCR, the area under the plasma concentration- europa.eu/docs/en_GB/document_library/Scientific_guideline/2012/07/ time curve (AUC) ratio, represents the fold increase in plasma AUC of WC500129606.pdf) is based on a similar type of ratio and cutoff value, a probe drug whose clearance is entirely determined by metabolism by namely, [I]unbound/Ki,unbound $ 0.02, where Ki,unbound is as defined earlier and [I]unbound is the unbound (free) maximum concentration of drug in plasma (mean Cmax obtained following treatment with the highest Parts of this work were previously presented at the following meeting: Haupt L, recommended clinical dose). The EMA’s guideline also applies to Kazmi F, Ogilvie B, Buckley D, Smith B, Leatherman S, and Parkinson A (2011) significant circulating metabolites, defined as phase 1 metabolites whose Can Ki values for direct inhibition of CYP enzymes be reliably estimated from IC50 . values? Seventeenth North American Regional ISSX Meeting; 2011 Oct 16–20; plasma AUC is 25% of the parent drug and 10% of total circulating Atlanta, GA. drug-derived material. To evaluate the potential for reversible inhibition dx.doi.org/10.1124/dmd.115.066597. of CYP3A enzymes in the intestine, the concentration of inhibitor in eq. 1 s This article has supplemental material available at dmd.aspetjournals.org. is defined as [I]gut = molar dose/250 ml. Values of [I]gut can greatly ABBREVIATIONS: AFE, average fold error; AUC, area under the plasma concentration-time curve; EMA, European Medicines Agency; FDA, U.S. Food and Drug Administration; NRMSE, normalized root mean square error; P450, cytochrome P450; RMSE, root mean square error. 1744 Estimating Ki from IC50 Values for P450 Inhibition 1745 exceed values of [I] (total Cmax,ss); accordingly, the FDA’s cutoff value for (internal standard), and dehydronifedipine-d6 (internal standard) were purchased inhibition of intestinal P450 enzymes is 11 (based on 1 + [I]gut/Ki,unbound). from SynFine Research (Richmond Hill, Ontario, Canada). 10-Deacetyltaxol (internal standard) was purchased from A.F. Hauser, Inc. Pharmaceutical The corresponding EMA cutoff is 10 (based on [I]gut/Ki,unbound). Typically, inhibition of P450 enzymes is first evaluated in vitro by (Valparaiso, IN). Investigational Drugs. determining the concentration of investigational drug (or significant A total of 132 investigational drugs and drug metabolites were examined during the course of P450 inhibition studies circulating metabolite) that causes 50% inhibition of P450 enzyme sponsored by numerous pharmaceutical companies in the United States, Europe, activity (IC50) with a selective P450 probe at a substrate concentration and Japan. Unfortunately, because they are proprietary compounds and for approximately equal to Km (Michaelis-Menten constant). Determining reasons of confidentiality, we are not at liberty to disclose the identity of the the mechanism of reversible inhibition (competitive, noncompetitive, structures of these compounds. The compounds represent a set of structurally mixed, or uncompetitive) and measuring the value of Ki requires an in diverse, small drug molecules under development for several different thera- vitro evaluation of the effects of multiple inhibitor concentrations peutic indications. The design of the experiments and the interpretation of the versus multiple substrate concentrations, ideally with the former results of this study (a comparison of two endpoints of P450 inhibition) required neither knowledge of chemical structures nor physicochemical properties. spanning Ki and the latter spanning Km. Determining values of IC50 and Test System. Pooled human liver microsomes (n = 16 or 200; mixed gender) Ki based on the unbound concentration of inhibitor in the test system (typically human liver microsomes) requires knowledge of fu (the were prepared from nontransplantable livers and characterized at XenoTech, inc LLC (Lenexa, KS) as described previously (Pearce et al., 1996; Parkinson et al., fraction of unbound drug in the microsomal incubation), which can be 2004). determined experimentally or estimated theoretically from the inhib- Incubation Conditions. K and IC values were determined in accordance Downloaded from ’ i 50 itor s logP or logD value and the concentration of microsomal protein with recommendations in the FDA and EMA guidance documents and consensus (Austin et al.., 2002; Hallifax and Houston, 2006). papers (Tucker et al., 2001; Bjornsson et al., 2003). All experiments were The relationship between Ki and values of IC50 determined when [S] performed under the following conditions: 1) the concentration of P450 marker (substrate concentration) = Km depends on the mechanism of inhibition, substrate was approximately equal to Km for IC50 determinations and spanned as summarized in Table 1 (Cheng and Prusoff, 1973; Brandt et al., Km for Ki determinations (i.e., [S] ranged from 0.25 times Km to 10 times Km, solubility permitting); 2) the substrate incubation time was 5 minutes to minimize 1987; Cer et al., 2009). In the case of noncompetitive inhibition, Ki = metabolism-dependent inhibition and inhibitor depletion; and 3) the concentra- dmd.aspetjournals.org IC50. In the case of competitive and uncompetitive inhibition, Ki =IC50/ 2. In the case of mixed inhibition, K values range from IC to IC /2. tion of human liver microsomes was 0.1 mg/ml or less to maximize the unbound i 50 50 inhibitor concentration. The FDA’s 2012 Guidance for Industry on drug interactions acknowl- In general, incubations were conducted at 37°C in 200- or 400-ml incubation edges that Ki values are often estimated from values of IC50/2, based on mixtures containing potassium phosphate buffer (50 mM, pH 7.4), MgCl2 the conservative assumption that the mechanism of reversible inhibition (3 mM), EDTA (1 mM), NADPH-generating system, and human liver microsomes.
Load more

Recommended publications
  • Hypochlorous Acid Handling
    Hypochlorous Acid Handling 1 Identification of Petitioned Substance 2 Chemical Names: Hypochlorous acid, CAS Numbers: 7790-92-3 3 hypochloric(I) acid, chloranol, 4 hydroxidochlorine 10 Other Codes: European Community 11 Number-22757, IUPAC-Hypochlorous acid 5 Other Name: Hydrogen hypochlorite, 6 Chlorine hydroxide List other codes: PubChem CID 24341 7 Trade Names: Bleach, Sodium hypochlorite, InChI Key: QWPPOHNGKGFGJK- 8 Calcium hypochlorite, Sterilox, hypochlorite, UHFFFAOYSA-N 9 NVC-10 UNII: 712K4CDC10 12 Summary of Petitioned Use 13 A petition has been received from a stakeholder requesting that hypochlorous acid (also referred 14 to as electrolyzed water (EW)) be added to the list of synthetic substances allowed for use in 15 organic production and handling (7 CFR §§ 205.600-606). Specifically, the petition concerns the 16 formation of hypochlorous acid at the anode of an electrolysis apparatus designed for its 17 production from a brine solution. This active ingredient is aqueous hypochlorous acid which acts 18 as an oxidizing agent. The petitioner plans use hypochlorous acid as a sanitizer and antimicrobial 19 agent for the production and handling of organic products. The petition also requests to resolve a 20 difference in interpretation of allowed substances for chlorine materials on the National List of 21 Allowed and Prohibited Substances that contain the active ingredient hypochlorous acid (NOP- 22 PM 14-3 Electrolyzed water). 23 The NOP has issued NOP 5026 “Guidance, the use of Chlorine Materials in Organic Production 24 and Handling.” This guidance document clarifies the use of chlorine materials in organic 25 production and handling to align the National List with the November, 1995 NOSB 26 recommendation on chlorine materials which read: 27 “Allowed for disinfecting and sanitizing food contact surfaces.
    [Show full text]
  • Assessment of Combined Toxicity of Heavy Metals from Industrial Wastewaters on Photobacterium Phosphoreum T3S
    Appl Water Sci DOI 10.1007/s13201-016-0385-4 ORIGINAL ARTICLE Assessment of combined toxicity of heavy metals from industrial wastewaters on Photobacterium phosphoreum T3S 1,2 1 2 1 2 BibiSaima Zeb • Zheng Ping • Qaisar Mahmood • Qiu Lin • Arshid Pervez • 2 2 2 Muhammad Irshad • Muhammad Bilal • Zulfiqar Ahmad Bhatti • Shahida Shaheen2 Received: 19 May 2015 / Accepted: 19 January 2016 Ó The Author(s) 2016. This article is published with open access at Springerlink.com Abstract This research work is focusing on the toxicities Introduction of heavy metals of industrial origin to anaerobic digestion of the industrial wastewater. Photobacterium phospho- The presence of heavy metals in excess amounts inter- reum T3S was used as an indicator organism. The acute feres with the beneficial uses of water because of the toxicities of heavy metals on P. phosphoreum T3S were toxicity of heavy metals and the biomagnification effect assessed during 15-min half inhibitory concentration brought on by its accumulation on ecology (Chang et al. (IC50) as indicator at pH 5.5–6. Toxicity assays involved 2006; Altas 2009). During recent times, heavy metals the assessment of multicomponent mixtures using TU and were focus of attention owing to their hazardous nature MTI approaches. The results of individual toxicity indi- and subsequent toxicity studies. Various workers have cated that the toxicity of Cd, Cu and Pb on P. phospho- assessed the combined toxicity of heavy metals (Su reum increased with increasing concentrations and there et al. 2012;Quetal.2013; Mochida et al. 2006). It has was a linear correlation. The 15-min IC50 values of Cd, Cu been argued that the effects caused by combinations of and Pb were 0.537, 1.905 and 1.231 mg/L, respectively, various heavy metals may be more threatening and and their toxic order was Cd [ Pb [ Cu.
    [Show full text]
  • Introduction
    Guidance for Assay Development & HTS March 2007 Version 5 Section I: Introduction Introduction Copyright © 2005, Eli Lilly and Company and the National Institutes of Health Chemical Genomics Center. All Rights Reserved. For more information, please review the Privacy Policy and Site Usage and Agreement. Table of Contents A. INTRODUCTION This document is written to provide guidance to investigators that are interested in developing assays useful for the evaluation of compound collections to identify chemical probes that modulate the activity of biological targets. Originally written as a guide for therapeutic projects teams within a major pharmaceutical company, this manual has been adapted to provide guidelines for: a. Identifying potential assay formats compatible with High Throughput Screen (HTS), and Structure Activity Relationship (SAR) b. Developing optimal assay reagents c. Optimizing assay protocol with respect to sensitivity, dynamic range, signal intensity and stability d. Adopting screening assays to automation and scale up in microtiter plate formats e. Statistical validation of the assay performance parameters f. Secondary follow up assays for chemical probe validation and SAR refinement g. Data standards to be followed in reporting screening and SAR assay results. General definition of biological assays This manual is intended to provide guidance in the area of biological assay development, screening and compound evaluation. In this regard an assay is defined by a set of reagents that produce a detectable signal allowing a biological process to be quantified. In general, the quality of an assay is defined by the robustness and reproducibility of this signal in the absence of any test compounds or in the presence of inactive compounds.
    [Show full text]
  • An Online Tool for Calculating and Mining Dose–Response Data Nicholas A
    Clark et al. BMC Cancer (2017) 17:698 DOI 10.1186/s12885-017-3689-3 SOFTWARE Open Access GRcalculator: an online tool for calculating and mining dose–response data Nicholas A. Clark1†, Marc Hafner2†, Michal Kouril3, Elizabeth H. Williams2, Jeremy L. Muhlich2, Marcin Pilarczyk1, Mario Niepel2, Peter K. Sorger2 and Mario Medvedovic1* Abstract Background: Quantifying the response of cell lines to drugs or other perturbagens is the cornerstone of pre-clinical drug development and pharmacogenomics as well as a means to study factors that contribute to sensitivity and resistance. In dividing cells, traditional metrics derived from dose–response curves such as IC50, AUC, and Emax, are confounded by the number of cell divisions taking place during the assay, which varies widely for biological and experimental reasons. Hafner et al. (Nat Meth 13:521–627, 2016) recently proposed an alternative way to quantify drug response, normalized growth rate (GR) inhibition, that is robust to such confounders. Adoption of the GR method is expected to improve the reproducibility of dose–response assays and the reliability of pharmacogenomic associations (Hafner et al. 500–502, 2017). Results: We describe here an interactive website (www.grcalculator.org) for calculation, analysis, and visualization of dose–response data using the GR approach and for comparison of GR and traditional metrics. Data can be user-supplied or derived from published datasets. The web tools are implemented in the form of three integrated Shiny applications (grcalculator, grbrowser, and grtutorial) deployed through a Shiny server. Intuitive graphical user interfaces (GUIs) allow for interactive analysis and visualization of data. The Shiny applications make use of two R packages (shinyLi and GRmetrics) specifically developed for this purpose.
    [Show full text]
  • Lack of Correlation Between in Vitro and in Vivo Studies on the Inhibitory Effects Of
    pharmaceutics Article Lack of Correlation between In Vitro and In Vivo Studies on the Inhibitory Effects of (-)-Sophoranone on CYP2C9 Is Attributable to Low Oral Absorption and Extensive Plasma Protein Binding of (-)-Sophoranone 1, 2,3, 3 3 3 Yu Fen Zheng y, Soo Hyeon Bae y , Zhouchi Huang , Soon Uk Chae , Seong Jun Jo , Hyung Joon Shim 3, Chae Bin Lee 3, Doyun Kim 3,4, Hunseung Yoo 4 and Soo Kyung Bae 3,* 1 School of Basic Medicine and Clinical Pharmacy, China Pharmaceutical University, 639 Longmian Road, Jiangning District, Nanjing 211198, China; [email protected] 2 Q-fitter, Inc., Seoul 06578, Korea; sh.bae@qfitter.com 3 College of Pharmacy and Integrated Research Institute of Pharmaceutical Sciences, The Catholic University, Korea, Bucheon 14662, Korea; [email protected] (Z.H.); [email protected] (S.U.C.); [email protected] (S.J.J.); [email protected] (H.J.S.); [email protected] (C.B.L.); [email protected] (D.K.) 4 Life Science R&D Center, SK Chemicals, 310 Pangyo-ro, Sungnam 13494, Korea; [email protected] * Correspondence: [email protected]; Tel.: +82-2-2164-4054 These authors contributed equally to this work. y Received: 8 March 2020; Accepted: 5 April 2020; Published: 7 April 2020 Abstract: (-)-Sophoranone (SPN) is a bioactive component of Sophora tonkinensis with various pharmacological activities. This study aims to evaluate its in vitro and in vivo inhibitory potential against the nine major CYP enzymes. Of the nine tested CYPs, it exerted the strongest inhibitory effect on CYP2C9-mediated tolbutamide 4-hydroxylation with the lowest IC50 (Ki) value of 0.966 0.149 µM (0.503 0.0383 µM), in a competitive manner.
    [Show full text]
  • (2R,6R)-Hydroxynorketamine Do Not Block NMDA Receptor Function
    Antidepressant-relevant concentrations of the ketamine metabolite (2R,6R)-hydroxynorketamine do not block NMDA receptor function Eric W. Lumsdena,1, Timothy A. Troppolib,1, Scott J. Myersc, Panos Zanosd, Yasco Aracavaa, Jan Kehre,f, Jacqueline Lovettg, Sukhan Kimc, Fu-Hua Wange,f, Staffan Schmidte,f, Carleigh E. Jenned, Peixiong Yuanh, Patrick J. Morrisi, Craig J. Thomasi, Carlos A. Zarate Jr.h, Ruin Moaddelg, Stephen F. Traynelisc,2, Edna F. R. Pereiraa,j,2, Scott M. Thompsonb,d,2, Edson X. Albuquerquea,j,k,2, and Todd D. Gouldd,j,l,m,2,3 aDepartment of Epidemiology and Public Health, Division of Translational Toxicology, University of Maryland School of Medicine, Baltimore, MD 21201; bDepartment of Physiology, University of Maryland School of Medicine, Baltimore, MD 21201; cDepartment of Pharmacology, Emory University, Atlanta, GA 30329; dDepartment of Psychiatry, University of Maryland School of Medicine, Baltimore, MD 21201; eDepartment of Physiology and Pharmacology, Karolinska Institute, SE-171 77 Stockholm, Sweden; fPronexus Analytical AB, SE-167 33 Bromma, Sweden; gBiomedical Research Center, National Institute on Aging, Intramural Research Program, National Institutes of Health, Baltimore, MD 21224; hSection on the Neurobiology and Treatment of Mood Disorders, Intramural Research Program, National Institute of Mental Health, National Institutes of Health, Bethesda, MD 20892; iDivision of Preclinical Innovation, National Center for Advancing Translational Sciences, Intramural Research Program, National Institutes of Health, Bethesda, MD 20892; jDepartment of Pharmacology, University of Maryland School of Medicine, Baltimore, MD 21201; kDepartment of Medicine, University of Maryland School of Medicine, Baltimore, MD 21201; lDepartment of Anatomy and Neurobiology, University of Maryland School of Medicine, Baltimore, MD 21201; and mVeterans Affairs Maryland Health Care System, Baltimore, MD 21201 Edited by Solomon H.
    [Show full text]
  • Glossary of Hazardous Materials Terms
    GLOSSARY OF CHEMICAL HAZARD TERMS ACGIH - The American Conference of Governmental Industrial Hygienists consists of occupational safety and health professionals who recommend occupational exposure limits for many substances. Action Level - An OSHA concentration calculated as an 8-your time-weighted average, which initiates certain required activities such as exposure monitoring and medical surveillance. Acute Health Effect - A severe effect which occurs rapidly after a brief intense exposure to a substance. ANSI - American National Standards Institute is a private group that develops consensus standards. Acute toxicity -Acutely toxic substances cause adverse effects by any of the following exposure methods: 1. Oral or dermal administration of a single dose of a substance. 2. Multiple oral or dermal doses within a 24-hour period 3. An inhalation exposure of 4 hours. Asphyxiant - A chemical (gas or vapor) that can cause death or unconsciousness by suffocation. Aspiration hazard - A liquid or solid chemical that causes severe acute effects if it infiltrates into the trachea and lower respiratory tract. Possible effects include chemical pneumonia, pulmonary injury, or death Autoignition Temperature - The lowest temperature at which a substance will burst into flames without a source of ignition like a spark or flame. The lower the ignition temperature, the more likely the substance is going to be a fire hazard. Boiling Point - The temperature of a liquid at which its vapor pressure is equal to the gas pressure over it. With added energy, all of the liquid could become vapor. Boiling occurs when the liquid's vapor pressure is just higher than the pressure over it. Carcinogen - A substance that causes cancer in humans or, because it has produced cancer in animals, is considered capable of causing cancer in humans.
    [Show full text]
  • Analyzing Binding Data UNIT 7.5 Harvey J
    Analyzing Binding Data UNIT 7.5 Harvey J. Motulsky1 and Richard R. Neubig2 1GraphPad Software, La Jolla, California 2University of Michigan, Ann Arbor, Michigan ABSTRACT Measuring the rate and extent of radioligand binding provides information on the number of binding sites, and their afÞnity and accessibility of these binding sites for various drugs. This unit explains how to design and analyze such experiments. Curr. Protoc. Neurosci. 52:7.5.1-7.5.65. C 2010 by John Wiley & Sons, Inc. Keywords: binding r radioligand r radioligand binding r Scatchard plot r r r r r r receptor binding competitive binding curve IC50 Kd Bmax nonlinear regression r curve Þtting r ßuorescence INTRODUCTION A radioligand is a radioactively labeled drug that can associate with a receptor, trans- porter, enzyme, or any protein of interest. The term ligand derives from the Latin word ligo, which means to bind or tie. Measuring the rate and extent of binding provides information on the number, afÞnity, and accessibility of these binding sites for various drugs. While physiological or biochemical measurements of tissue responses to drugs can prove the existence of receptors, only ligand binding studies (or possibly quantitative immunochemical studies) can determine the actual receptor concentration. Radioligand binding experiments are easy to perform, and provide useful data in many Þelds. For example, radioligand binding studies are used to: 1. Study receptor regulation, for example during development, in diseases, or in response to a drug treatment. 2. Discover new drugs by screening for compounds that compete with high afÞnity for radioligand binding to a particular receptor.
    [Show full text]
  • Synthesis and Cytotoxicity Evaluation of Novel Asymmetrical Mono-Carbonyl Analogs of Curcumin (Amacs) Against Vero, Hela, and MCF7 Cell Lines
    Scientia Pharmaceutica Article Synthesis and Cytotoxicity Evaluation of Novel Asymmetrical Mono-Carbonyl Analogs of Curcumin (AMACs) against Vero, HeLa, and MCF7 Cell Lines Pekik Wiji Prasetyaningrum, Anton Bahtiar ID and Hayun Hayun * ID Faculty of Pharmacy, Universitas Indonesia, Depok 16424, West Java, Indonesia; [email protected] (P.W.P.); [email protected] (A.B.) * Correspondence: [email protected]; Tel.: +62-21-727-0031 Received: 8 May 2018; Accepted: 5 June 2018; Published: 7 June 2018 Abstract: A series of novel asymmetrical mono-carbonyl analogs of curcumin (AMACs) were synthesized and evaluated for cytotoxic activity using BSLT and MTT assay against Vero, HeLa, and MCF7 cell lines. The structures of the synthesized compounds were confirmed by FTIR, 1H-NMR, 13C-NMR, and mass spectral data. The results of the cytotoxicity evaluation showed that the synthesized compounds exhibited moderate to very high toxic activity in BSLT (LC50 value 29.80–1704.23 µM); most of the compound exhibited cytotoxic activity against HeLa cell lines, which is comparable to the activity of cisplatin (IC50 value 40.65–95.55 µM), and most of the compound tested against MCF7 cell lines exhibited moderate to very high cytotoxic activity (IC50 value 7.86–35.88 µM). However, the selectivity index (SI) of the compounds was low (<1–1.96). Among the synthesized compounds, compound 1b was the most cytotoxic and selective against MCF7 cell lines. It could be considered for further development to obtain the more active and selective chemotherapeutic agents against breast cancer. Keywords: asymmetrical mono-carbonyl analogs of curcumin; AMACs; synthesis; cytotoxicity; Vero; HeLa; MCF7; cell lines 1.
    [Show full text]
  • International Union of Pharmacology Committee on Receptor Nomenclature and Drug Classification
    0031-6997/03/5504-597–606$7.00 PHARMACOLOGICAL REVIEWS Vol. 55, No. 4 Copyright © 2003 by The American Society for Pharmacology and Experimental Therapeutics 30404/1114803 Pharmacol Rev 55:597–606, 2003 Printed in U.S.A International Union of Pharmacology Committee on Receptor Nomenclature and Drug Classification. XXXVIII. Update on Terms and Symbols in Quantitative Pharmacology RICHARD R. NEUBIG, MICHAEL SPEDDING, TERRY KENAKIN, AND ARTHUR CHRISTOPOULOS Department of Pharmacology, University of Michigan, Ann Arbor, Michigan (R.R.N.); Institute de Recherches Internationales Servier, Neuilly sur Seine, France (M.S.); Systems Research, GlaxoSmithKline Research and Development, Research Triangle Park, North Carolina (T.K.); and Department of Pharmacology, University of Melbourne, Parkville, Australia (A.C.) Abstract ............................................................................... 597 I. Introduction............................................................................ 597 II. Working definition of a receptor .......................................................... 598 III. Use of drugs in definition of receptors or of signaling pathways ............................. 598 A. The expression of amount of drug: concentration and dose ............................... 598 1. Concentration..................................................................... 598 2. Dose. ............................................................................ 598 B. General terms used to describe drug action ...........................................
    [Show full text]
  • Neurotoxicity Screening of New Psychoactive Substances (NPS): Effects on Neuronal Activity in Rat Cortical Cultures Using Microelectrode Arrays (MEA)
    Accepted Manuscript Title: Neurotoxicity screening of new psychoactive substances (NPS): effects on neuronal activity in rat cortical cultures using microelectrode arrays (MEA) Authors: Anne Zwartsen, Laura Hondebrink, Remco HS Westerink PII: S0161-813X(18)30086-X DOI: https://doi.org/10.1016/j.neuro.2018.03.007 Reference: NEUTOX 2313 To appear in: NEUTOX Received date: 25-10-2017 Revised date: 9-3-2018 Accepted date: 19-3-2018 Please cite this article as: Zwartsen A, Hondebrink L, Westerink RH, Neurotoxicity screening of new psychoactive substances (NPS): effects on neuronal activity in rat cortical cultures using microelectrode arrays (MEA), Neurotoxicology (2010), https://doi.org/10.1016/j.neuro.2018.03.007 This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. Neurotoxicity screening of new psychoactive substances (NPS): effects on neuronal activity in rat cortical cultures using microelectrode arrays (MEA) Anne Zwartsen a,b, Laura Hondebrink b, Remco HS Westerink a,* a Neurotoxicology Research Group, Division Toxicology, Institute for Risk Assessment Sciences (IRAS), Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands. b Dutch Poisons Information Center (DPIC), University Medical Center Utrecht, Utrecht University, Utrecht, The Netherlands. * Corresponding author: Dr. Remco HS Westerink. Neurotoxicology Research Group, Division Toxicology, Institute for Risk Assessment Sciences (IRAS), Faculty of Veterinary Medicine, Utrecht University, P.O.
    [Show full text]
  • First Principles and Their Application to Drug Discovery
    REVIEWS Drug Discovery Today Volume 17, Numbers 1/2 January 2012 The utilization of the kinetic and thermodynamic signatures of preclinical leads is proving pivotal in their triage and rational optimization towards clinical candidates with maximal in vivo efficacy devoid of adverse events. Reviews KEYNOTE REVIEW Target–drug interactions: first principles and their application to drug discovery 1 1 Sara Nu´n˜ez studied organic Sara Nu´n˜ ez , Jennifer Venhorst and Chris G. Kruse chemistry at the University of Barcelona (Spain) and the Abbott Healthcare Products, 1381 CP Weesp, The Netherlands University of London (UK). She received her Ph.D. in 2003 from the University of Manchester (UK), and thereafter did a In this review, we begin by introducing the basic principles of kinetics postdoc in Biophysics at the and thermodynamics of target–drug binding within the context of Albert Einstein College of Medicine (USA). In 2005, she drug discovery. In addition, we present a meta-analysis of the recent joined Solvay Pharmaceuticals (now Abbott Healthcare) in The Netherlands as a postdoctoral fellow; and in 2008, she literature describing the kinetic and thermodynamic resolution of was promoted to Sr. Computational Medicinal Chemist. At Abbott, she has supported the medicinal chemistry efforts successful clinical candidates with diverse mechanisms of action. for neuroscience drug discovery programs, from target We finish by discussing the best practices in the triage and chemical discovery up to and including clinical proof of principle studies. She has supported more than 15 programs optimization towards clinical candidates with maximal in vivo internationally, and was project manager of the D2-103 Top Institute Pharma innitiative.
    [Show full text]